波谱分析课件 chapter 5 MS.pptVIP

Chapter 5Mass Spectrometry Mass Spectrometry Basics What information can be determined? Molecular weight Molecular formula (HRMS) Structure (from fragmentation fingerprint) Isotopic incorporation / distribution Protein sequence (MS-MS) Mass Spectrometry Basics Mass spectrometry has 4 basic operations: Sample introduction (analyte must be in vapor phase) Ionization Mass analysis ( separating ions by mass/charge ratio) Detection and quantitation Sample Introduction Method Applications Batch (reservoir) gases, volatile liquids Direct insertion probe very low vapor pressure solids and liquids Membrane aqueous solutions, air samples Chromatography eluent LC-MS, GC-MS, etc. Ionization Methods 1. Electron Ionization (EI) most common ionization technique, limited to relatively low MW compounds (600 amu) 2. Chemical Ionization (CI) ionization with very little fragmentation, still for low MW compounds (800 amu) 3. Desorption Ionization (DI) for higher MW or very labile compounds 4. Spray ionization (SI) for LC-MS, biomolecules, etc. Electron Ionization (EI) vaporized sample is bombarded with high energy electrons (typically 70 eV) “hard” ionization method leads to significant fragmentation ionization is efficient but non-selective PLK Chapter 8 is written from an EIMS perspective Electron Ionization Advantages inexpensive, versatile and reproducible fragmentation gives structural information large databases if EI spectra exist and are searchable Disadvantages fragmentation at expense of molecular ion sample must be relatively volatile Chemical Ionization (CI) Vaporized sample reacts with pre-ionized reagent gas via proton transfer, charge exchange, electron capture, adduct formation, etc. common CI reagents: methane, ammonia, isobutane, hydrogen, methanol “soft” ionization gives little fragmentation selective ionization--only exothermic or thermoneutral ion-molecule reactions will occur choice of reagent allows tuning of ionizati